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Implications of Male Amelogenin Dropouts in Forensics

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Implications of Male Amelogenin Dropouts in Forensics Research Article J Forensic Sci & Criminal Inves Volume 15 Issue 2 - February 2021 Copyright © All rights are reserved by Anand Kumar DOI: 10.19080/JFSCI.2021.15.555908 Implications of Male Amelogenin Dropouts in Forensics Anand Kumar* DNA Division, State Forensic Science Laboratory, India Submission: February 06, 2021; Published: February 22, 2021 *Corresponding author: Anand Kumar, DNA Division, State Forensic Science Laboratory, Rajasthan, 302016, (INDIA) Abstract In forensic science STR loci are useful tools for reconstructing male lineages, paternity testing, forensic investigations, and population The samples used in this study were received at State Forensic Science Laboratory for routine examination. A combination of autosomal (Power Plexgenomics.®- Fusion They 5C cover system a widekit) and range Y-STR of genome-specific(Power Plex®- Y23 geographical system kit) distributions multiplexing duesystems to shortfall was used of forrecombination the genotyping during of the spermatogenesis. samples as per the manufacturer’s protocol. In electropherogram of male samples, male-derived amelogenin marker was not observed in the results of Power Plex®- Fusion 5C system kit. These samples were further analyzed by Power Plex®- Y23 system kit and a complete set of 23 Y STR markers was obtained. The failure rate of detection of amelogenin marker in Indian population is 0.23%. This clearly indicates the deletion in the short arm of Y chromosome related to amelogenin marker. Keywords: Amelogenin marker; Forensic genetics; Haplogroups; Deletion Introduction DNA analysis based on autosomal as well as sex chromosome STR is routinely used in forensic casework, population studies, identification [6], Thangaraj et al [7], reported 1.85% failure in Southern hybridization [5]. Some studies reported that the primer medico-legal examinations etc. [1-4]. In humans, sex-determining the amplification of AMELY locus in 270 Indian males through binding site region includes point mutation instead of AMELY genes known as amelogenins are present on both the X and the Y mutation [8,9]. Interstitial deletion of AMELY locus at short arm chromosomes. A single-copy gene (Amelogenin gene) located on of Y chromosome was also reported [10]. AMELY deletion in Xp22.1eXp22.3 and Yp11.2 was sequenced by [5]. Length of AMELY various populations has also been reported by several workers and AMELX genes is 3272 bp and 2872 bp respectively. It has been viz., in Australian [11], and Italian [12] populations, where Haas- X amelogenin gene should always be present in the reaction observed that a single reaction amplifies both genes, and therefore, primer producing PCR fragments using a different method for which offers a positive control. Amelogenin gene codes for tooth Rocholz and Weiler demonstrated smaller flanking sequences in the determination of gender in null AMELY males [13]. Lack of Y-amelogenin in 6 out of 29,432 (0.02%)Austrian males has also deletion due to the presence of its counterpart X chromosome, enamel. Enamel formation is not significantly affected by AMELY but it helps in forensic interpretation. Difference in size of these companies are manufacturing various multiplexing systems chromosomes has been recognized in forensic casework. It offers a been demonstrated [14]. For the purpose of gender identification, that incorporate amelogenin marker. Our study using Power remarkable tool for distinguishing between the evidence of victim Plex®- Fusion 5C system kit inferred a deletion at the short arm and offender in sexual assault cases. As a result of AMELY deletion, of Y chromosome (amelogenin gene) in the male population of normal males may be observed as female because of failure in the Rajasthan. It was determined by using PowerPlex®- Y23 system kit. lead to errors in forensic inference. Various studies have been amplification of the gene. Misinterpretation of the marker may performed showing mutation in AMELY genes. This mutation may Materials and Methods lead to serious consequences, if the males are typed as females due to AMELY deletion particularly in cases of rape or human The samples for the study were taken from the routine casework received at DNA division State Forensic Science Laboratory, Jaipur J Forensic Sci & Criminal Inves 15(2): JFSCI.MS.ID.555908 (2021) 001 Journal of Forensic Sciences & Criminal Investigation with prior written informed consent from the candidates as per Result and Discussion the declaration of Helsinki and were subjected to DNA isolation with autosomal markers in a single reaction, it is used in USA-Thermo) on Automate Express system (Thermo) according Since X and Y amelogenin markers can be amplified along by using Prep Filer Express TM kit (Thermo Fisher Scientific, CA, of the electropherogram of male samples demonstrated a forensic casework analysis for gender identification. Analysis to the recommended protocol of the manufacturer. Quantification of isolated samples were performed by using Quanti filer Trio ®- Fusion 5C dropout of Y-specific amelogenin (AMEL Y) marker because of kit (Thermo) on Quant Studio 5 system (Thermo). Amplification multiplexing system (Figure 1). This was further reinforced by system kit for 22 autosomal STRs viz., D3S1358, D2S441, vWA, amplification of one additional Y specific marker DYS391 in the of 1ng quantified DNA was done using Power Plex D21S11, D10S1248, D13S317, PENTA-E, D16S539, THO1, D18S51, of Y-chromosome with the help of PowerPlex®- Y23 system D2S1338, TPOX, CSF1PO, PENTA-D, D7S820, D5S818, D8S1179, successful amplification of 23 loci dispersed along both arms kit. In this manner, our study established the presence of AMEL Ydropouts in the population of Rajasthan which is in concordance ®- Y23 D12S391, D19S433, FGA, D1S1656andD22S1045. Amplification with outcomes of similar studies on Indian population [15]. system kit for 23 Y STRs like Y_GATA_ H4, DYS389I, DYS385a/b, of Y specific markers was carried out by using Power Plex Dropout in AMELY shows pinpoint deletion and the possibility of DYS389II, DYS391, DYS481, DYS392, DYS549, DYS533, DYS438, subsidiary chromosomal aberration needs to be explored. Dropout DYS437, DYS570, DYS635, DYS390, DYS439, DYS643, DYS393, of AMELY marker encourages the integration of additional Y DYS458, DYS448, DYS19, DYS456 and DYS576according to the recommended protocol except for half reaction volume. PCR more useful in the populations where AMELY dropout frequency products were subjected to Genetic Analyzer 3500 XL using chromosome specific markers in the multiplexing system. It is is very high. Although frequency of amelogenin dropout is very POPTM-4 (Performance Optimized Polymer), 36cm Capillary low (1.85%) in population of Rajasthan, conclusions should not be drawn based on this marker alone. The routinely used Y array. Each amplified sample was mixed with 9.5 µL Hi-Di done at 1.2 kV for 5s. Electrophoresis results were analyzed with Formamide and 0.5 µL ILS 500. Injection of the samples was loci simultaneously, ranging between 239 to 263 bp and 353-385 GeneMapper ID-X v1.6 software (Thermo). specific marker DYS389 is most suitable because it explores two bp respectively [16]. Figure 1: AMELY Dropout and Amplification of DYS391 locus in the sample. How to cite this article: Kertzman S, Kagan A, Vainder M, Hegedish O, Lapidus R, Weizman, A. Narcissistic Personality Measures Discriminate Between 002 Young Women With and Without Tattoos. J Forensic Sci & Criminal Inves. 2021; 15(2): 555906 DOI: 10.19080/JFSCI.2021.15.555906 Journal of Forensic Sciences & Criminal Investigation Acknowledgment Int J Legal Med 116(2): 121-123. Authors is grateful to the Director, State Forensic Science for gender identification in forensic casework and prenatal diagnosis. 8. Roffey PE, Eckhoff CI, Kuhl JL (2001) A rare mutation in the amelogenin Laboratory, Jaipur, Rajasthan for encouraging the study. 1016-1019. gene and its potential investigative ramifications. J Forensic Sci 45(5): Compliance with Ethical Standards 9. Henke J, Henke L, Chatthopadhyay P, Kayser M, Dulmer M, et al. (2001) As per the declaration of Helsinki, written informed consent Application of Y-chromosomal STR haplotypes to forensic genetics. Croat Med J 42(3): 292-297. was obtained for the study. 10. Lattanzi W, Di Giacomo MC, Lenato GM (2005) A large interstitial References deletion encompassing the amelogenin gene on the short arm of the Y chromosome. Hum Genet 116(5): 395-401. 1. Tandem Repeat (STR) in forensic science to Crackdown complex cases 11. Mitchell RJ, Kreskas M, Baxter E, Buffalino L, Van Oorschot RAH (2006) ofKumar sexual A, abuses. Kumar Adv R, MohsinBiores 10(5): U, Sharma 29-34. S. Genetic Profiling of Short An investigation of sequence deletions of amelogenin (AMELY), a Y-chromosome locus commonly used for gender determination. Ann 2. Kumar A, Kumar R, Kumawat RK, Tilawat A, Shrivastava P, et al. (2020) Hum Biol 33(2): 227-240. Genetic variation (population database) at 20 autosomal STR loci in the population of Rajasthan (north-western India). Int J Legal Med 12. Turrina S, Filippini G, Voglino G, De Leo D (2011) Two additional 134(5): 1-3. reports of deletion on the short arm of the Y chromosome. Forensic Sci Int Genet 5(3): 242-246. 3. Kumar A, Kumar R, Kumawat RK (2020) Genetic portrait study for 23 Y-STR loci in the population of Rajasthan, India. Int J Legal Med 134(5): 13. Haas Rochholz H, Weiler G (1997) Additional primer sets for an 1691-1693. amelogenin gene PCR-based DNA-sex test. Int J Legal Med 110(6): 312-315. 4. Anand Kumar, Rajesh Kumar, R. K. Kumawat, Ginni Kumawat (2021) Maternal allele mutation: Slippage synthesis furnishing evolutionary 14. Steinlechner M, Berger B, Niederstätter H, Parson W (2002) Rare trend. GSC Biological and Pharmaceutical Sciences 14(01): 090-094. failures in the amelogenin sex test. Int J Legal Med 116(2): 117-120. 5. 15. Kashyap VK, Sahoo S, Sitalaximi T, Trivedi R (2006) Deletions in the by polymerase chain reaction using X-Y homologous primer. Am J Med Y-derived amelogenin gene fragment in the Indian population.
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